title: Identification of Host Kinase Genes Required for Influenza Virus Replication and the Regulatory Role of MicroRNAs.
abstract: Human protein kinases (HPKs) have profound effects on cellular responses. To better understand the role of HPKs and the signaling networks that influence influenza virus replication, a small interfering RNA (siRNA) screen of 720 HPKs was performed. From the screen, 17 HPKs (NPR2, MAP3K1, DYRK3, EPHA6, TPK1, PDK2, EXOSC10, NEK8, PLK4, SGK3, NEK3, PANK4, ITPKB, CDC2L5 (CDK13), CALM2, PKN3, and HK2) were validated as essential for A/WSN/33 influenza virus replication, and 6 HPKs (CDK13, HK2, NEK8, PANK4, PLK4 and SGK3) were identified as vital for both A/WSN/33 and A/New Caledonia/20/99 influenza virus replication. These HPKs were found to affect multiple host pathways and regulated by miRNAs induced during infection. Using a panel of miRNA agonists and antagonists, miR-149* was found to regulate NEK8 expression, miR-548d-3p was found to regulate MAPK1 transcript expression, and miRs -1228 and -138 to regulate CDK13 expression. Up-regulation of miR-34c induced PLK4 transcript and protein expression and enhanced influenza virus replication, while miR-34c inhibition reduced viral replication. These findings identify HPKs important for influenza viral replication and show the miRNAs that govern their expression.
authors.affiliation: Department of Infectious Diseases, University of Georgia, Athens, Georgia, United States of America.
authors.forename: Abhijeet
authors.initials: A
authors.lastname: Bakre
authors.forename: Lauren E
authors.initials: LE
authors.lastname: Andersen
authors.forename: Victoria
authors.initials: V
authors.lastname: Meliopoulos
authors.forename: Keegan
authors.initials: K
authors.lastname: Coleman
authors.forename: Xiuzhen
authors.initials: X
authors.lastname: Yan
authors.forename: Paula
authors.initials: P
authors.lastname: Brooks
authors.forename: Jackelyn
authors.initials: J
authors.lastname: Crabtree
authors.forename: S Mark
authors.initials: SM
authors.lastname: Tompkins
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Tripp
mesh_terms: D000072283:A549 Cells; D000818:Animals; D001483:Base Sequence; D004285:Dogs; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D007251:Influenza, Human / Q000201:enzymology / Q000235:genetics* / Q000473:pathology / Q000821:virology*; D048728:MAP Kinase Kinase Kinase 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D061985:Madin Darby Canine Kidney Cells; D035683:MicroRNAs / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000072379:NIMA-Related Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D019590:Nucleocapsid Proteins; D010641:Phenotype; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012337:RNA, Ribosomal, 18S / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D016415:Sequence Alignment; D015854:Up-Regulation; D014758:Viral Core Proteins / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics*
chemical_list: D035683:MicroRNAs; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D012337:RNA, Ribosomal, 18S; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins; D011494:Protein Kinases; C490219:PLK4 protein, human; C458529:NEK8 protein, human; D000072379:NIMA-Related Kinases; D017346:Protein Serine-Threonine Kinases; D048728:MAP Kinase Kinase Kinase 1